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Avastin防治青光眼滤过术后瘢痕化的实验研究

发布时间:2018-11-03 07:02
【摘要】: 目的:利用兔眼青光眼滤过术(小梁切除)模型,予滤过泡旁球结膜下注射三种不同剂量的Avastin,探讨Avastin在抑制青光眼滤过术后瘢痕化方面的疗效,结膜下注射的时机和剂量,同时对Avastin结膜下注射是否对术区组织产生毒性反应作初步研究。 方法:眼压正常的新西兰白兔50只50眼随机分为三个不同浓度的Avastin实验组,即0.5mgAvastin实验组(A组)、1.0mgAvastin实验组(B组)、2.0mgAvastin实验组(C组),以及丝裂霉素C(MMC)对照组和空白对照组,每组10眼。对兔眼行常规小梁切除术,A、B、C实验组于术毕及术后第3、7天滤过泡旁球结膜下分别注射6.25mg/m1、12.50mg/ml、25.0mg/ml Avastin 0.08ml,即0.5、1.0、2.0mg;MMC对照组术中筋膜囊、巩膜瓣下放置浸湿0.2mg/m1 MMC棉片;空白对照组于术毕及术后第3、7天滤过泡旁球结膜下注射0.9%氯化钠注射液0.08ml。术后裂隙灯显微镜观察滤过泡情况,并行活体共焦显微镜检查;测量眼压;于术后第7、14天,每组摘除眼球5只,常规HE染色,于高倍镜下计算滤过泡腔面及形成区面积占结膜面积的平均百分比,Tunel法检测细胞凋亡情况,同时ELISA法测定房水及组织匀浆TGF-β1,bFGF含量。 结果:(1)裂隙灯显微镜检查:术后第7天,A、B、C实验组及MMC对照组滤过泡隆起弥散,表面血管稀疏;空白对照组术后第3天滤过泡开始局限化,至第7天滤过泡周围怒张粗大的血管形成。术后第14天,C实验组及MMC对照组滤过泡扁平弥散,表面血管稀疏纤细;B实验组滤过泡扁平弥散,表面血管较7天时密集;A实验组滤过泡血管包裹局限;空白对照组滤过泡瘢痕化、表面血管浓密。(2)眼压:术后14天内各组眼压均随时间呈线性增长,但增长趋势无差别,且不同时间点各组间眼压无差异。(3)活体共焦显微镜检查:术后第7天,A、B、C实验组及MMC对照组较空白对照组滤过泡囊腔结构明显,纤维组织疏松;MMC对照组组织层间见大量炎症细胞浸润。术后第14天,C实验组及MMC对照组仍可见明显滤过泡囊腔结构,纤维组织纤细疏松;B实验组残留少许滤过泡结构;其余各组纤维组织致密,纵横交错,血管粗大呈螺旋状。各组角膜结构均未见异常。(4)HE染色:术后第7天,各组可见明显滤过泡结构。术后第14天,C实验组及MMC对照组可见明显滤过泡结构;B实验组见少量微小的滤过泡;A实验组及空白对照组滤过泡消失,纤维组织增生明显。采用单因素方差分析显示:术后第7天,各组滤过泡腔面及形成区面积与结膜面积的平均百分比无差别;术后第14天,C实验组及MMC对照组滤过泡腔面占结膜的平均面积百分比高于A、B实验组及空白对照组,B实验组滤过泡腔面在结膜的平均面积百分比高于A实验组及空白对照组,而C实验组与MMC对照组间及A实验组与空白对照组间面积百分比无差别。C实验组及MMC对照组滤过泡形成区占结膜平均面积百分比高于A、B实验组及空白对照组;C实验组与MMC对照组间及A、B实验组与空白对照组间无差别。(5)ELISA法测定TGF-β1、bFGF含量:术后第7天及第14天,C实验组及MMC对照组房水、术区组织中TGF-β1、bFGF含量均低于A、B实验组及空白对照组;B实验组TGF-β1、bFGF含量低于A实验组及空白对照组;C实验组与MMC对照组组间及A实验组与空白对照组间TGF-β1、bFGF含量无差别。(6)Tunel细胞凋亡检测:术后第7天,各实验组及空白对照组术区组织中偶见强荧光的凋亡细胞,MMC对照组见大量凋亡细胞。术后第14天,A、B实验组及空白对照组偶见凋亡细胞;C实验组结膜上皮下、筋膜囊组织见少量凋亡细胞;MMC对照组仍见大量凋亡细胞。采用单因素方差分析显示:术后第7、14天,MMC对照组凋亡细胞均最多,高于各实验组及空白对照组;空白对照组与实验组组向凋亡细胞数无差别 结论:术后早期滤过泡旁球结膜下注射2.0mg及1.0mgAvastin可有效抑制滤过泡新生血管的形成,有助于缓解滤过泡的瘢痕化,促进功能性滤过泡的形成与维持,且安全性较好。
[Abstract]:Objective: To study the therapeutic effect of Avastin in the treatment of glaucoma after filtration, and to explore the time and dosage of avastin in the treatment of glaucoma after filtration. At the same time, the toxicity of Avastin subconjunctival injection was studied. Methods: 50 eyes of New Zealand white rabbits with normal intraocular pressure were randomly divided into three experimental groups with different concentrations, namely 0. 5mgAvastin experimental group (group A), 1. 0mgAvastin experimental group (group B), 2. 0mgAvastin experimental group (group C), and MMC (MMC) control group and blank control group. 10 eyes of group A, B and C were respectively injected with 6. 25mg/ ml, 12.50mg/ ml, 25. 0mg/ ml Avastin 0. 08ml, that is, 0. 5, 1. 0, 2. 0mg. MC cotton tablets, blank control group were injected with 0.9% sodium chloride injection at the 3 rd and 7 th day after operation, and 0.9% sodium chloride injection was injected. 08ml. After operation, the slit lamp microscope was used to observe the filter bubble condition, and the confocal microscopy was performed in parallel; the intraocular pressure was measured; after 7 and 14 days after operation, 5 eyes were removed from each group, conventional HE staining was performed, and the filtration bubble cavity surface and the area occupied by the formation area accounted for the level of the conjunctival area at high magnification. Both percentage and Tunel method were used to detect the apoptosis of the cells. GF content. Results: (1) Slit lamp microscope examination: After 7 days after operation, the alveolar ridge was dispersed in group A, B, C and MMC control group, and the surface blood vessel was sparse; 3 days after operation of blank control group, the bleb began to localize until the 7th day after filtration bubble. During the 14th day after operation, the experimental group and MMC control group filtered the flat dispersion, the surface blood vessels were sparse and thin, the experimental group was filtered with flat dispersion, and the surface vessels were dense at 7 days. (2) Intraocular pressure: The intraocular pressure in each group increased linearly with time within 14 days after operation, but there was no difference in the growth trend and different time. There was no difference in intraocular pressure in each group. (3) In-vivo confocal microscope examination: group A, B, C and MMC in control group were compared with blank control group, the structure of alveolar cavity was obvious, the fibrous tissue was loose, and the tissue layers of MMC control group A lot of inflammatory cell infiltration was found. After the 14th day of operation, the experimental group and MMC control group still showed a clear filter vesicle cavity structure, and the fibrous tissue was fine and loose; B experimental group had a little residual filtration cell structure; the other groups of fibrous tissue were dense and dry. and the blood vessels are in a spiral shape. The corneal structure was abnormal. (4) HE staining: 7 days after operation, each group The results showed that the bleb structure was found in the experimental group and the MMC control group after the 14th day of operation. The experimental group showed a small amount of small filtration bubble; the experimental group and the blank control group were filtered out to disappear. The results of single-factor analysis of variance showed that there was no difference between the surface area of each group and the area of formation area and the average percentage of conjunctival area after 7 days of operation. The percentage of soaking area was higher than that of group A, group B and blank control group, and the percentage of average area of filter cell in group B was higher than that of group A and blank control group, and between control group and MMC control group and control group A and control group B. The percentage of interconjunctival area in experimental group and MMC control group was higher than that of group A, group B and blank control group, and the experimental group and MMC control group and group A and B were compared with control group. There was no difference in the blank control group. (5) The content of bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, The content of bFGF in experimental group was lower than that of group A and blank control group, and between experimental group and MMC control group and between group A and blank control group. There was no difference in bFGF content. (6) Apoptosis of Tunel cells: In the 7th day after operation, the apoptotic cells of strong fluorescence were occasionally observed in each experimental group and the blank control group. In group A, group B and blank control group, apoptotic cells were occasionally seen in group A, group B and blank control group after operation. A large number of apoptotic cells were still seen in the group. The single factor analysis of variance showed that the apoptotic cells in the MMC control group were most in the 7th and 14th days after operation, higher than those in each experimental group and the control group, and the blank control group and the experimental group. group's orientation There was no difference in the number of apoptotic cells: 2. 0mg and 1. 0mgAvasin could effectively inhibit the formation of filter bubble neovascularization at the early stage after operation.
【学位授予单位】:暨南大学
【学位级别】:硕士
【学位授予年份】:2010
【分类号】:R779.6

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